The objective of this work is the determination of the metabolic pathways for the conversion of 4-dimethylallyltryptophan (DMAT) to the clavine alkaloids and to other products and the characterization of the enzymes involved in these reactions. DMAT is converted to clavicipitic acid in cell-free systems from Claviceps sp. The enzyme DMAT oxidase (forming clavicipitic acid) will be purified. Spectrophotometric and chemical studies of the prosthetic group and kinetic studies will be carried out. The possible involvement of superoxide ion and the presence of reaction intermediates will be investigated. Chemical synthesis of intermediates in clavine alkaloid biosynthesis between DMAT and chanoclavine I will be synthesized via N- tosyl indolyl-4-acetaldehyde. 4-(Z-4-hydroxy-3-methyl- delta squared- butenyl)- tryptophan (Z-HODMAT) will be synthesized by a known procedure. Radioactive or unlabeled methyl iodide will be used in the synthesis of N alpha-methyl derivatives of E- and Z-HODMAT and DMAT. The epoxide and allylic alcohol derivatives of DMAT will be synthesized. The radioactive compounds will be tested for incorporation into the clavine alkaloids in cultures of Claviceps sp. Compounds which are incorporated will be studied further. The presence of the compound in cultures will be investigated. The formation of the compound from other substrates and the conversion of the compound to products in cell-free systems will be investigated. If efficient conversion in cell-free extracts is obtained, the enzyme involved will be studied. The synthesized compounds will be tested as substrates or inhibitors of DMAT oxidase (forming clavicipitic acid).